【摘要】 为了研究3个不同蛋白质水平(10%、18%、26%)饲粮对肥育猪的钙蛋白抑制蛋白酶(CAST)的影响,对CAST蛋白提取、电泳、转膜等条件进行优化。试验表明:CAST被激活的条件是将蛋白提取液于上样缓冲液95℃煮15min后,立即在冰块上冷却,防止蛋白降解;采用分离胶浓度为10%进行电泳,可以明显检测到68kD、107 kD的CAST;采用预染Marker和DAB染色液可针对性地检测到目的蛋白的位置;利用上述程序检测到了猪CAST两种类型的表达,取得了较满意的结果。更多还原

【Abstract】 Western blotting method to investigate effects of different dietary levels protein(10%,18% and 26%) on the expression of CAST of growing-finishing pigs was studied.Its procedures including the extraction of protein,the condition of electrophoresis and immuno-blotting were optimized.The results showed that CAST was activated after the extract was heated at 95℃ for 15min and then cooled on the ice immediately in order to prevent being degraded.CAST of 68 kD and 107 kD was detected by using a separation gel of 10% for SDS-PAGE preparation,the target protein position was detected directly by pre-stained Marker and DAB staining solution.Two types of CAST were detected successfully by the above procedures.更多还原